Journal: PLoS ONE
Article Title: Prdm8 Regulates the Morphological Transition at Multipolar Phase during Neocortical Development
doi: 10.1371/journal.pone.0086356
Figure Lengend Snippet: In utero electroporation of any one of control (pCAG-IRES-EGFP with pCAG-IRES-Puro; A), or Prdm8 gain-of-function (pCAG-IRES-EGFP with pCAG-Prdm8; B), or Prdm8 loss-of-function (pCAG-IRES-EGFP with pPrdm8sh#629; C) vectors were carried out at E12.5, and the brains were analyzed at P5. The cortex was divided into 10 bins and the percentage of EGFP-positive cells were quantified (D). The distribution of EGFP-positive cells is significantly increased in the upper bins (bins8–10) and reduced in lower bins (bin5) in the case of the pCAG-Prdm8-electroporated brains, and significantly decreased in upper bins (bins4–6), and increased in lower bins (bins1, 2) in the pPrdm8sh-electroporated brains. The number of counted cells was about 137 cells. Data represents the mean ± SD (n = 6 slices from 3 individuals); *p<0.05, **p<0.01. High-power images showing that the molecular features of control, Prdm8 gain-of-function, or loss-of-function cells in the neocortex, stained with Tbr1 (E, F, G), Ctip2 (K, L, M), RORb (K, L, M) and Brn2 (H, I, J). The percentage of each layer marker-positive EGFP-positive cells located in each layer position was quantified (N). The number of counted cells was about 184 cells. Data represents the mean ± SD (n>4 slices from 4 individuals); **p<0.01. Scale bars: 100 µm (A,E).
Article Snippet: The antibodies used were, rat anti-GFP (1∶500; nakalai tesk), rabbit anti-GFP (1∶200; IBL), rabbit anti-DsRed (1∶500; Invitrogen), goat anti-Unc5D (1∶100; R&D), rabbit anti-Tbr2 (1∶300; abcam), goat anti-NeuroD1 (1∶100; Santa Cruz), mouse anti-PCNA (1∶100; Cell Signaling), mouse anti-Tuj1 (1∶500; SIGMA), rabbit anti-Tbr1 (1∶100; abcam), mouse anti-RORb (1∶100; PERSEUS PROTEOMICS), rat anti-Ctip2 (1∶300; abcam), goat anti-Brn2 (1∶100; Santa Cruz), and mouse anti-Prdm8 .
Techniques: In Utero, Electroporation, Staining, Marker
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